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Offlinetrip_hazard
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First time playing with agar - so far, bupkis
    #26575446 - 04/03/20 03:41 PM (3 years, 9 months ago)

After reading every link on the Agar Portal and getting my shit together I finally landed on Munchauzen's agar tech.  I poured and PC'd a dozen containers and decided to try my hand at cleaning up two strains - PE from syringe and AA+ from spore.  I did my best to follow clean procedure in my SAB and did the zig-zag swipe for both; that was on 3/30.  I have checked on these buggers every day and so far I've got nada - no bacteria, no myc, no random contaminants, just nothing.  Is it possible to screw up the 123 agar method so badly that I can't even contaminate a plate?





Edited by trip_hazard (04/03/20 06:26 PM)


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OfflineLogicaL ChaosM
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Re: First time playing with agar - so far, bupkis [Re: trip_hazard]
    #26575493 - 04/03/20 04:02 PM (3 years, 9 months ago)

How long did u let the agar cool after PCing?


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Offlinetrip_hazard
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Re: First time playing with agar - so far, bupkis [Re: LogicaL Chaos]
    #26575501 - 04/03/20 04:04 PM (3 years, 9 months ago)

Two days - got busy and just left the PC on the stove closed for a couple days before I had a chance to noc'em up.


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InvisibleMunchauzen
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Re: First time playing with agar - so far, bupkis [Re: trip_hazard]
    #26576027 - 04/03/20 08:03 PM (3 years, 9 months ago)

spores can take up to two weeks to germinate. give it a few more days.


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Invisiblebrindle foxx
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Re: First time playing with agar - so far, bupkis [Re: Munchauzen]
    #26576037 - 04/03/20 08:05 PM (3 years, 9 months ago)

Quote:

Munchauzen said:
spores can take up to two weeks to germinate. give it a few more days.



I seem to not be able to grow spores from print on Agar maybe I need to put them in a syringe solution. Tissue seems to be the only thing to work for me.. maybe i need to stop using premix PDA also I seem to let the agar cool until its chunky and the pour is ugly


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OfflineBibBonk
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Re: First time playing with agar - so far, bupkis [Re: trip_hazard]
    #26576100 - 04/03/20 08:28 PM (3 years, 9 months ago)

Quote:

Is it possible to screw up the 123 agar method so badly that I can't even contaminate a plate?




I don't think so.

It's only been 4 days, that's not long enough to declare them failed.

If you want faster germination you can incubate them at 80F.


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Offlinetrip_hazard
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Re: First time playing with agar - so far, bupkis [Re: Munchauzen]
    #26576185 - 04/03/20 09:16 PM (3 years, 9 months ago)

Quote:

Munchauzen said:
spores can take up to two weeks to germinate. give it a few more days.




:thumbup:

Thanks - I think I've read too many threads where they're seeing partial coverage after a few days.  What's been your longest duration to visual confirmation?


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Offlinetrip_hazard
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Re: First time playing with agar - so far, bupkis [Re: BibBonk]
    #26576210 - 04/03/20 09:28 PM (3 years, 9 months ago)

Quote:

BibBonk said:
Quote:

Is it possible to screw up the 123 agar method so badly that I can't even contaminate a plate?




I don't think so.

It's only been 4 days, that's not long enough to declare them failed.

If you want faster germination you can incubate them at 80F.




I've been out of the game for awhile - I'll submit it's possible that I've forgotten the first rule.  Patience.


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